Abstract：
:Next-generation sequencing technology is a powerful tool for transcriptome analysis. However, under certain conditions, only a small amount of material is available, which requires more sensitive techniques that can preferably be used at the single-cell level. Here we describe a single-cell digital gene expression profiling assay. Using our mRNA-Seq assay with only a single mouse blastomere, we detected the expression of 75% (5,270) more genes than microarray techniques and identified 1,753 previously unknown splice junctions called by at least 5 reads. Moreover, 8-19% of the genes with multiple known transcript isoforms expressed at least two isoforms in the same blastomere or oocyte, which unambiguously demonstrated the complexity of the transcript variants at whole-genome scale in individual cells. Finally, for Dicer1(-/-) and Ago2(-/-) (Eif2c2(-/-)) oocytes, we found that 1,696 and 1,553 genes, respectively, were abnormally upregulated compared to wild-type controls, with 619 genes in common.
journal_name
Nat Methodsjournal_title
Nature methodsauthors
Tang F,Barbacioru C,Wang Y,Nordman E,Lee C,Xu N,Wang X,Bodeau J,Tuch BB,Siddiqui A,Lao K,Surani MAdoi
10.1038/nmeth.1315subject
Has Abstractpub_date
2009-05-01 00:00:00pages
377-82issue
5eissn
1548-7091issn
1548-7105pii
nmeth.1315journal_volume
6pub_type
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